USPTO Patents Application 10551504

Applicant
Serial No.
Filed
Page

Hiroyuki Tsunoda et al.

10/551,504

May 12, 2006

2

Attorney's Docket No. 14875-0153US1

Amendments to the Claims :

This listing of claims replaces all prior versions and listings of claims in the application:
Listing of Claims :

1 . (Previously presented) An antibody comprising a single-chain polypeptide that binds
to the thrombopoietin (TPO) receptor (Mpl), wherein said antibody comprises two heavy chain
variable regions and two light chain variable regions, and at least one of the heavy chain variable
regions comprises a set of CDR1, CDR2 and CDR3 sequences selected from the group
consisting of:

(a) SEQ ID NOs: 27, 28, and 29;

(b) SEQ ID NOs: 36, 37, and 38; and

(c) SEQ ID NOs: 57, 58, and 59.

2. (Previously presented) The antibody of claim 1, wherein the two heavy chain variable
regions and the two light chain variable regions are arranged in the order, from the N-terminus of
the single-chain polypeptide, of heavy chain variable region, light chain variable region, heavy
chain variable region, and light chain variable region.

3. (Previously presented) The antibody of claim 1, wherein the four variable regions are
linked by linkers.

4. (Previously presented) The antibody of claim 3, wherein each linker comprises 15
amino acids.

5. (Canceled)

Applicant : Hiroyuki Tsunoda et al. Attorney's Docket No. 14875-0153US1

Serial No. : 10/551,504

Filed : May 12, 2006

Page : 3

6. (Previously presented) The antibody of claim 1, which is a humanized antibody.

7. (Canceled)

8. (Previously presented) The antibody of claim 1 , wherein the antibody binds to soluble

Mpl.

9. -10. (Canceled)

1 1 . (Previously presented) The antibody of claim 1 , wherein the antibody binds to
soluble Mpl with a KD = 10" 6 M or lower.

12. (Previously presented) The antibody of claim 1, wherein the antibody binds to
soluble Mpl with a KD = 10" 7 M or lower.

13. (Previously presented) The antibody of claim 1, wherein the antibody has a TPO
agonistic activity of EC50 = 100 nM or lower.

14. (Previously presented) The antibody of claim 1, wherein the antibody has a TPO
agonistic activity of EC50 = 30 nM or lower.

15. (Previously presented) The antibody of claim 1, wherein the antibody has a TPO
agonistic activity of EC50 = 10 nM or lower.

16. (Canceled)

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10/551,504

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Attorney's Docket No. 14875-0153US1

17. (Previously presented) The antibody of claim 1, wherein at least one of the light
chain variable regions comprises a set of CDR1, CDR2 and CDR3 sequences selected from the
group consisting of:

(a) SEQ ID NOs: 84, 85, and 86;

(b) SEQ ID NOs: 93, 94, and 95; and

(c) SEQ ID NOs: 114, 115, and 116.

1 8. (Currently amended) The antibody of claim 1 comprising any one of:

(a) heavy chain variable region CDR1, CDR2, and CDR3 comprising amino acid
sequences consisting of SEQ ID NOs: 27, 28, and 29, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising amino acid sequences consisting of SEQ ID NOs:
84, 85, and 86, respectively;

(b) heavy chain variable region CDR1, CDR2, and CDR3 comprising amino acid
sequences consisting of SEQ ID NOs: 36, 37, and 38, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising amino acid sequences consisting of SEQ ID NOs:
93, 94, and 95, respectively; and

(c) heavy chain variable region CDR1, CDR2, and CDR3 comprising amino acid
sequences consisting of SEQ ED NOs: 9, 10, and 1 1 57, 58 and 59 , respectively, and light chain
variable region CDR1, CDR2, and CDR3 comprising amino acid sequences consisting of SEQ
ID NOs: 114, 115, and 116, respectively.

19. -22. (Canceled)

23. (Previously presented) The antibody of claim 1, wherein at least one of the heavy
chain variable regions comprises a set of FR1, FR2, FR3, and FR4 sequences selected from the
group consisting of:

(a) SEQ ID NOs: 230, 232, 234, and 236; and

(b) SEQ ID NOs: 265, 267, 269, and 271.

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Hiroyuki Tsunoda et al.

10/551,504

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Attorney's Docket No. 14875-0153US1

24. (Previously presented) The antibody of claim 1, wherein at least one of the light
chain variable regions comprises a set of FR1, FR2, FR3, and FR4 sequences selected from the
group consisting of:

(a) SEQ ID NOs: 239, 241, 243, and 245; and

(b) SEQ ID NOs: 272, 274, 276, and 278.

25. (Previously presented) The antibody of claim 1, comprising any one of:

(a) heavy chain variable region FR1, FR2, FR3, and FR4 comprising amino acid
sequences consisting of SEQ ID NOs: 230, 232, 234, and 236, respectively, and light chain
variable region FR1, FR2, FR3, and FR4 comprising amino acid sequences consisting of SEQ
ID NOs: 239, 241, 243, and 245, respectively; and

(b) heavy chain variable region FR1, FR2, FR3, and FR4 comprising amino acid
sequences consisting of SEQ ID NOs: 265, 267, 269, and 271, respectively, and light chain
variable region FR1, FR2, FR3, and FR4 comprising amino acid sequences consisting of SEQ ID
NOs: 272, 274, 276, and 278, respectively.

26. (Previously presented) The antibody of claim 1, wherein at least one of said heavy
chain variable regions comprises the amino acid sequence of SEQ ID NO: 140, 162, or 229.

27. (Previously presented) The antibody of claim 1, wherein at least one of said light
chain variable regions comprises the amino acid sequence of SEQ ID NO: 141, 163, or 238.

28. (Previously presented) The antibody of claim 1 , wherein:

(a) at least one of the heavy chain variable regions comprises the amino acid sequence of
SEQ ID NO: 229, and at least one of the light chain variable regions comprises the amino acid
sequence of SEQ ID NO: 238;

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10/551,504

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Attorney's Docket No. 14875-0153US1

(b) at least one of the heavy chain variable regions comprises the amino acid sequence of
SEQ ID NO: 140, and at least one of the light chain variable regions comprises the amino acid
sequence of SEQ ID NO: 141; or

(c) at least one of the heavy chain variable regions comprises the amino acid sequence of
SEQ ID NO: 162, and at least one of the light chain variable regions comprises the amino acid
sequence of SEQ ID NO: 163.

29.-31. (Canceled)

32. (Previously presented) The antibody of claim 1, wherein the antibody recognizes an
epitope within the region of amino acids 26 to 274 of human Mpl (SEQ ID NO: 123).

33. (Previously presented) The antibody of claim 1, which has TPO agonistic activity.

34. -37. (Canceled)

38. (Previously presented) A pharmaceutical composition comprising the antibody of
claim 32.

39. -43. (Canceled)

44. (Previously presented) A method of treating or reducing the incidence of
thrombocytopenia in a subject in need thereof, comprising administering to the subject a
therapeutically effective amount of the pharmaceutical composition of claim 38.

45. (Previously presented) A method for increasing platelet number in a subject in need
thereof, comprising administering to the subject a therapeutically effective amount of the
pharmaceutical composition of claim 38.

Applicant: Hiroyuki Tsunoda et al. Attorney's Docket No. 14875-0153US1

Serial No. : 10/551,504

Filed : May 12, 2006

Page : 7

46. (Previously presented) The method of claim 45, wherein the pharmaceutical
composition is administered to the subject after the administration of platelet components to the
subject.

47. (Previously presented) A method for increasing the amount of platelet components at
the time of blood collection, the method comprising preadministering to a blood collection
subject an effective amount of the pharmaceutical composition of claim 38.

48. (Previously presented) The method of claim 47, further comprising collecting blood
from the subject after said preadministration.